Rehan Mujeeb Faridi

Influence of activating and inhibitory killer immunoglobulin-like receptors on predisposition to recurrent miscarriages

BACKGROUND Understanding of the immune events and mechanisms occurring at the feto-maternal interface is likely to help in understanding the ability of the fetus to survive within the maternal body. Evidence supporting extensive roles of natural killer cells during pregnancy gives rise to a possibility that these NK cells can be mis-regulated and involved in fetal allograft rejection. Killer immunoglobulin-like receptors (KIR) play an important role in regulating the NK cell activity through their activating and inhibiting isoforms. Since there exists a considerable, genetically determined variation in the repertoire of KIR receptors between different individuals, a particular maternal KIR repertoire may predispose to recurrent miscarriages (RMs). METHODS Gene-specific PCR amplification (PCR-SSP) was used to determine the individual KIR genotypes in women experiencing RM and controls. RESULTS A higher prevalence of activating KIR genes was seen in patients than in controls. Among women experiencing RM, the BB genotypes were more prevalent (P < 0.0001, OR = 4.4, 95% CI = 2.89-6.69) compared with controls. CONCLUSIONS Our results indicate that the balance between inhibitory and activating receptor-mediated signals present in natural killer cells is inclined toward a more activating state that may contribute to pregnancy loss.

Genetic Association of Phase I and Phase II Detoxification Genes with Recurrent Miscarriages among North Indian Women

Allelic variants of the detoxification genes that have impaired biotransformation functions may increase susceptibility to reproductive toxicity leading to endometriosis, recurrent miscarriage (RM) or poor pregnancy outcome. In the present study, we have investigated CYP1A1, CYP2D6, GSTT1, GSTP1 and GSTM1, which are involved in the phase I and phase II detoxification systems, in relation to their role in the etiology of unexplained RMs. In a case-control study, we have investigated 200 females with RM and 300 age and ethnically matched healthy controls with successful reproductive history from North India. The frequencies of phase I wild-type genotypes of CYP1A1 and CYP2D6 in RM cases were 0.56 and 0.60, whereas in controls these were 0.68 and 0.65, respectively (both P < 0.05). The GSTM1 null-genotype frequencies were 0.66 and 0.84 among RM cases and controls, respectively, the GSTT1 null-genotype frequencies were 0.52 and 0.45 (P < 0.005) and the GSTP1 variant allele frequencies were 0.23 and 0.20, respectively. In conclusion, we observed significant protective effects of phase I wild-type genotypes and association of the GSTT1 null genotype with RM. Through combined analyses we have highlighted the importance of the balance of phase I/phase II detoxification systems, in the etiology of RM.

Vascular endothelial growth factor gene polymorphisms in North Indian patients with recurrent miscarriages

The association of four common polymorphisms of vascular endothelial growth factors (VEGF) with recurrent miscarriages(RM) was evaluated in North Indian women for 200 patients with RM and 200 controls. The subjects were genotyped for the polymorphisms 2578C/A, 2549 18-bp I/D, 1154G/A and +936C/T. Association of VEGF genotypes, alleles and haplotypes with recurrent miscarriage were evaluated by Fisher’s exact test. 1154G/A and +936C/T modified the risk of RM. The 1154A allel and +936T allel significantly increased the risk of RM (OR = 1.485, P = 0.0210, 95% CI 1.072–2.057 and OR = 1.869, P = 0.0054, 95% CI 1.214–2.876 respectively). Risk was further increased when –1154A/A genotype and +936C/T genotype were considered (OR = 2.0, P = 0.0310,95% CI 1.068–3.747 and OR = 1.716, P = 0.0293, 95% CI 1.058–2.784 respectively). However, no association was found between 2578C/A or 2549 18-bp I/D and RM. Four haplotypes, AIAC, ADAC, CIAT and ADGT, were found to predispose to RM while the haplotypes CIAC, CDGT and ADGC were found to show protective effect. In conclusion, two common polymorphisms of the VEGF gene,1154G/A and +936C/T, increase the risk of RM in North Indian women. RM is also predisposed in the presence of haplotypes AIAC,ADAC, CIAT and ADGT.

Interleukin-1 gene cluster variants and recurrent pregnancy loss among North Indian women: retrospective study and meta-analysis

The aim of this study was to determine whether or not interleukin-1 alpha (IL-1a), interleukin-1 beta (IL-1b) and IL-1 receptor antagonist (IL-1RA) polymorphisms are associated with risk of unexplained recurrent pregnancy loss (RPL) among North Indian women. This retrospective case–control study examine 200 well-characterized RPL cases for IL-1 gene cluster variants, determined by restriction fragment length polymorphism-PCR. The observed allele, genotype and haplotype distributions were compared with those obtained from 300 ethnically matched negative controls. Invariant distribution of IL-1 gene cluster single-nucleotide polymorphisms was observed among RPL cases and controls. Meta-analysis of IL-1b _511, +3953 and IL-1RN 86-bp variable number tandem repeat from the reported literature and this study did not reveal any significant association with the risk of RPL. In conclusion, no significant difference between RPL and control groups was observed at the allele, genotype or haplotype levels when tested for association using the dominant, recessive and additive models of inheritance for IL-1 gene cluster variants. As far as is known, this is the first report from India pertaining to IL-1 gene cluster variants’ association with the risk of RPL from North India.

Upregulation of HLA-G in JEG-3 cells by dexamethasone and hydrocortisone.

BackgroundVarious reports suggest that HLA-G molecule plays an important role in feto-maternal interface, protecting the allogenic fetus from maternal immune attack. It is shown that steroid hormones may upregulate the HLA-G gene expression. In the present study, we have made an attempt to upregulate the HLA-G gene expression in a HLA-G+ve cell line (JEG-3) by using two glucocorticoids drugs, i.e., dexamethasone and hydrocortisone.MethodsChoriocarcinoma JEG-3 (HLA-G+ve), JAR (HLA-G−ve) and erythroleukemia K-562 (HLA-G−ve) cell lines were obtained from American Type Culture Collection. These cell lines were treated with glucocorticoids (dexamethasone and hydrocortisone). HLA-G gene transcription was determined by standard and real-time RT-PCR analysis, and protein expression was evaluated by both flow cytometry and Western blotting.ResultsDose-dependent increase in HLA-G mRNA and protein expression was observed in HLA-G+ve JEG-3 cells, while no expression was recorded in JAR and K-562 (HLA-G−ve) cell lines.ConclusionWe were able to upregulate HLA-G expression only in HLA-G+ve cell line. On the basis of our results, we hypothesize that the HLA-G gene expression can be upregulated only when the cell lines/cells have the basal expression and not in the cells that totally lack its expression. We have further hypothesized that these drugs may be used only in those women with recurrent miscarriages who show minimum basal expression level of HLA-G.

Genetic analysis of eNOS gene polymorphisms in association with recurrent miscarriage among North Indian women

This study investigated the association of common polymorphisms of the endothelial nitric oxide synthase (eNOS) gene with recurrent miscarriage (RM) among North Indian women. A total of 200 patients with unexplained recurrent miscarriages and 300 controls were genotyped for six polymorphic regions of eNOS by PCR, re-sequencing and RFLP. The GG genotype of 12862A>G, the G allele of Glu298Asp and the aa genotype of intron 4VNTR increased the risk of RM by ∼1.8-fold, ∼3.5-fold and ∼2-fold, respectively (odds ratio (OR) 1.84, 95% confidence intervals (CI) 1.19-2.86, P=0.0066; OR 3.58, 95% CI 2.12-6.03, P<0.0001; and OR 2.23, 95% CI 1.04-4.77, P=0.0493). Two haplotypes were found to have a significant protective effect against RM (OR 0.63, 95% CI 0.48-0.82, P=0.0009; and OR 0.4, 95% CI 0.19-0.81, P=0.0149) and another was found to increase the risk of RM by ∼2-fold (OR 2.12, 95% CI 1.16-3.89 P=0.0195). In conclusion three common polymorphisms of eNOS gene, 12862A>G, Glu298Asp and intron 4VNTR increase the risk of RM in North Indian women. Risk of RM may also be modified by the presence of particular haplotypes.

Recurrent pregnancy loss and apolipoprotein E gene polymorphisms: a case–control study from north India.

Citation Agarwal M, Parveen F, Faridi RM, Phadke SR, Das V, Agrawal S. Recurrent pregnancy loss and apolipoprotein E gene polymorphisms: a case–control study from North India. Am J Reprod Immunol 2010; 64: 172–178

Genetic association of adipokine and UCP2 polymorphism with recurrent miscarriage among non-obese women

The adipokines produced from adipose tissues influence energy homeostasis, resulting in alterations of the adipokine concentrations. This process may be associated with fertility impairment, resulting in recurrent miscarriage. The present study investigated whether there was any association between the UCP2 45-bp indel polymorphism and the adipokine gene polymorphisms, namely leptin 2549 (C/A), adeponectin 276 (G/T) and 45 (T/G) and resistin 420 (C/G) in 200 non-obese recurrent miscarriage patients and 300 ethnically matched negative controls. These markers were studied using gene-specific PCR single specific primer and restriction fragment length polymorphism. For leptin 2549 and adeponectin 276, the A allele and G allele showed 3.42-fold (P=0.0001) and 1.36-fold (P=0.036) increased risk of recurrent miscarriage, respectively. Combined analysis of UCP2 45-bp indel and leptin 2549 showed U0-L0 and U1-L0 variants to be at 2- and 3-fold increased associative risk, respectively. Combined analysis of leptin 2549 and adeponectin 276 showed L0-D0 and L0-D1 variants to be at 2- and 4-fold increased associative risk, respectively. The combination U1-L0-D1-A1-R1 was 4.39-fold higher (P=0.0007) among recurrent miscarriage patients. In conclusion, the results highlight the role of the studied adipokine and UCP2 polymorphisms in recurrent miscarriage among the North Indian non-obese population. Pregnancy invokes a large shift in maternal metabolism. The normal concentrations of adipokines, which maintain the integrity of the hypothalamus-pituitary-gonadal axis, regular ovulatory processes and successful embryo implantation, are altered because of the influence of energy homeostasis, which in turn leads to fertility impairment and recurrent miscarriage of unknown aetiology. Recurrent miscarriage is reported in higher frequency among obese women. The UCP2 45-bp indel polymorphism and the adipokine gene polymorphisms namely leptin 2549 (C/A), adeponectin 276 (G/T), adeponectin 45 (T/G) and resistin 420 (C/G) have been shown to be associated with obesity. Most of the adipokine-related studies done previously have taken into consideration the metabolic function and obesity. However, there exist very few studies to evaluate the role of adipokines in non-obese recurrent miscarriage with no cause of repeated pregnancy losses. The present study focused at evaluating the independent effect of these single-nucleotide polymorphisms in non-obese women undergoing recurrent miscarriage.

Analysis of CCR5 and CX3CR1 gene polymorphisms in association with unexplained recurrent miscarriages among north Indian women

CONTEXT Recurrent miscarriage (RM), defined as three or more consecutive losses before the 20th week of gestation, affects 0.5-2% of pregnant women. In over 80% of cases, RM remains unexplained after investigations, suggesting the involvement of genetic factors. OBJECTIVES The present study investigates the common polymorphisms of chemokine receptors CCR5 (NG_012637.1:g.5303A>G) and CX3CR1 (NG_016362.1:g.21065C>T, Thr280Met and NG_016362.1:g.20971G>A, Val249Ile) and their association with recurrent miscarriages (RM) among north Indian women. PARTICIPANTS AND METHODS In a retrospective case-control study 200 well characterized patients with unexplained RM and 300 controls were genotyped for three polymorphic markers of CCR5 and CX3CR1 by restriction digestion of PCR amplified fragments. RESULTS Alleles and genotypes of CX3CR1 Val249Ile revealed statistically significant associations with RM cases when compared with the controls. The homozygous variant genotype Ile/Ile was found to be significantly higher among patients (p=0.0002) when compared with the homozygous wild type Val/Val genotype. The haplotype of CX3CR1 that carried major alleles of Thr280Met and Val249Ile (T-V) showed statistically significant protective association (p<0.0001, OR=0.41, 95% CI=0.31-0.54). The haplotype A-T-V (all wild type alleles) revealed a statistically significant protective association (p<0.0001, OR=0.41, 95% CI=0.34-0.62), whereas the haplotypes G-T-I, A-T-I and A-M-V modified the risk of RM 1.9-fold, 5.5-fold and 5.1-fold respectively. CONCLUSIONS A common polymorphism of CX3CR1 gene, Val240Ile is associated with the risk of RM in north Indian women. Risk of RM may also be modified by the presence of haplotypes T-I, M-V, G-T-I, A-T-I and A-M-V.

Acetylcholinesterase gene polymorphism and recurrent pregnancy loss

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